Pyrrolo[3,4-d]pyrimidines as corticotropin releasing factor (CRF) antagonists

ABSTRACT

Corticotropin releasing factor (CRF) antagonists of Formula (II)and their use in treating anxiety, depression, and other psychiatric, neurological disorders as well as treatment of immunological, cardiovascular or heart-related diseases and colonic hypersensitivity associated with psychopathological disturbance and stress.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.09/898,326, filed Jul. 3, 2001, now allowed, which in turn claimspriority of provisional application Serial No. 60/216,887, filed Jul. 7,2000. Said prior applications are incorporated herein by reference intheir entirety, for all purposes.

FIELD OF THE INVENTION

This invention relates to a treatment of psychiatric disorders andneurological diseases including major depression, anxiety-relateddisorders, post-traumatic stress disorder, supranuclear palsy andfeeding disorders as well as treatment of immunological, cardiovascularor heart-related diseases and colonic hypersensitivity associated withpsychopathological disturbance and stress, by administration of certainpyrrolo[3,4-d]pyrimidines.

BACKGROUND OF THE INVENTION

Corticotropin releasing factor (herein referred to as CRF), a 41 aminoacid peptide, is the primary physiological regulator ofproopiomelanocortin (POMC)—derived peptide secretion from the anteriorpituitary gland [J. Rivier et al., Proc. Nat. Acad. Sci. (USA) 80:4851(1983); W. Vale et al., Science 213:1394 (1981)]. In addition to itsendocrine role at the pituitary gland, immunohistochemical localizationof CRF has demonstrated that the hormone has a broad extrahypothalamicdistribution in the central nervous system and produces a wide spectrumof autonomic, electrophysiological and behavioral effects consistentwith a neurotransmitter or neuromodulator role in brain [W. Vale et al.,Rec. Prog. Horm. Res. 39:245 (1983); G. F. Koob, Persp. Behav. Med. 2:39(1985); E. B. De Souza et al., J. Neurosci. 5:3189 (1985)]. There isalso evidence that CRF plays a significant role in integrating theresponse of the immune system to physiological, psychological, andimmunological stressors [J. E. Blalock, Physiological Reviews 69:1(1989); J. E. Morley, Life Sci. 41:527 (1987)].

Clinical data provide evidence that CRF has a role in psychiatricdisorders and neurological diseases including depression,anxiety-related disorders and feeding disorders. A role for CRF has alsobeen postulated in the etiology and pathophysiology of Alzheimer'sdisease, Parkinson's disease, Huntington's disease, progressivesupranuclear palsy and amyotrophic lateral sclerosis as they relate tothe dysfunction of CRF neurons in the central nervous system [for reviewsee E. B. De Souza, Hosp. Practice 23:59 (1988)].

In affective disorder, or major depression, the concentration of CRF issignificantly increased in the cerebral spinal fluid (CSF) of drug-freeindividuals [C. B. Nemeroff et al., Science 226:1342 (1984); C. M. Bankiet al., Am. J. Psychiatry 144:873 (1987); R. D. France et al., Biol.Psychiatry 28:86 (1988); M. Arato et al., Biol Psychiatry 25:355(1989)]. Furthermore, the density of CRF receptors is significantlydecreased in the frontal cortex of suicide victims, consistent with ahypersecretion of CRF [C. B. Nemeroff et al., Arch. Gen. Psychiatry45:577 (1988)]. In addition, there is a blunted adrenocorticotropin(ACTH) response to CRF (i.v. administered) observed in depressedpatients [P. W. Gold et al., Am J. Psychiatry 141:619 (1984); F.Holsboer et al., Psychoneuroendocrinology 9:147 (1984); P. W. Gold etal., New Eng. J. Med. 314:1129 (1986)]. Preclinical studies in rats andnon-human primates provide additional support for the hypothesis thathypersecretion of CRF may be involved in the symptoms seen in humandepression [R. M. Sapolsky, Arch. Gen. Psychiatry 46:1047 (1989)]. Thereis preliminary evidence that tricyclic antidepressants can alter CRFlevels and thus modulate the numbers of CRF receptors in brain[Grigoriadis et al., Neuropsychopharmacology 2:53 (1989)].

There has also been a role postulated for CRF in the etiology ofanxiety-related disorders. CRF produces anxiogenic effects in animalsand interactions between benzodiazepine/non-benzodiazepine anxiolyticsand CRF have been demonstrated in a variety of behavioral anxiety models[D. R. Britton et al., Life Sci. 31:363 (1982); C. W. Berridge and A. J.Dunn Regul. Peptides 16:83 (1986)]. Preliminary studies using theputative CRF receptor antagonist a-helical ovine CRF (9-41) in a varietyof behavioral paradigms demonstrate that the antagonist produces“anxiolytic-like” effects that are qualitatively similar to thebenzodiazepines [C. W. Berridge and A. J. Dunn Horm. Behav. 21:393(1987), Brain Research Reviews 15:71 (1990)]. Neurochemical, endocrineand receptor binding studies have all demonstrated interactions betweenCRF and benzodiazepine anxiolytics providing further evidence for theinvolvement of CRF in these disorders. Chlordiazepoxide attenuates the“anxiogenic” effects of CRF in both the conflict test [K. T. Britton etal., Psychopharmacology 86:170 (1985); K. T. Britton et al.,Psychopharmacology 94:306 (1988)] and in the acoustic startle test [N.R. Swerdlow et al., Psychopharmacology 88:147 (1986)] in rats. Thebenzodiazepine receptor antagonist (Ro15-1788), which was withoutbehavioral activity alone in the operant conflict test, reversed theeffects of CRF in a dose-dependent manner while the benzodiazepineinverse agonist (FG7142) enhanced the actions of CRF [K. T. Britton etal., Psychopharmacology 94:306 (1988)].

The mechanisms and sites of action through which the standardanxiolytics and antidepressants produce their therapeutic effects remainto be elucidated. It has been hypothesized however, that they areinvolved in the suppression of the CRF hypersecretion that is observedin these disorders. Of particular interest is that preliminary studiesexamining the effects of a CRF receptor antagonist α-helical CRF₉₋₄₁) ina variety of behavioral paradigms have demonstrated that the CRFantagonist produces “anxiolytic-like” effects qualitatively similar tothe benzodiazepines [for review see G. F. Koob and K. T. Britton, In:Corticotropin-Releasing Factor: Basic and Clinical Studies of aNeuropeptide, E. B. De Souza and C. B. Nemeroff eds., CRC Press p221(1990)].

Several publications describe corticotropin releasing factor antagonistcompounds and their use to treat psychiatric disorders and neurologicaldiseases. Examples of such publications include DuPont Merck PCTapplication US94/11050, Pfizer WO 95/33750, Pfizer WO 95/34563, PfizerWO 95/33727 and Pfizer EP 0778 277 A1.

Insofar as is known, [1,5-a]-pyrazolo-1,3,5-triazines,[1,5-a]-1,2,3-triazolo-1,3,5-triazines, [1,5-a]-pyrazolo-pyrimidines and[1,5-a]-1,2,3-triazolo-pyrimidines, have not been previously reported ascorticotropin releasing factor antagonist compounds useful in thetreatment of psychiatric disorders and neurological diseases. However,there have been publications which teach some of these compounds forother uses.

For instance, EP 0 269 859 (Ostuka, 1988) discloses pyrazolotriazinecompounds of the formula

where R¹ is OH or alkanoyl, R² is H, OH, or SH, and R³ is an unsaturatedheterocyclic group, naphthyl or substituted phenyl, and states that thecompounds have xanthine oxidase inhibitory activity and are useful fortreatment of gout.

EP 0 594 149 (Ostuka, 1994) discloses pyrazolotriazine andpyrazolopyrimidine compounds of the formula

where A is CH or N, R⁰ and R³ are H or alkyl, and R¹ and R² are H,alkyl, alkoxyl, alkylthio, nitro, etc., and states that the compoundsinhibit androgen and are useful in treatment of benign prostatichypertrophy and prostatic carcinoma.

U.S. Pat. No. 3,910,907 (ICI, 1975) discloses pyrazolotriazines of theformula:

where R1 is CH₃, C₂H₅ or C₆H₅, X is H, C₆H₅, m-CH₃C₆H₄, CN, COOEt, Cl, Ior Br, Y is H, C₆H₅, o-CH₃C₆H₄, or p-CH₃C₆H₄, and Z is OH, H, CH₃, C₂H₅,C₆H₅, n-C₃H₇, i-C₃H₇, SH, SCH₃, NHC₄H₉, or N(C₂H₅)₂, and states that thecompounds are c-AMP phosphodiesterase inhibitors useful asbronchodilators.

U.S. Pat. No. 3,995,039 discloses pyrazolotriazines of the formula:

where R1 is H or alkyl, R² is H or alkyl, R³ is H, alkyl, alkanoyl,carbamoyl, or lower alkylcarbamoyl, and R is pyridyl, pyrimidinyl, orpyrazinyl, and states that the compounds are useful as bronchodilators.

U.S. Pat. No. 5,137,887 discloses pyrazolotriazines of the formula

where R is lower alkoxy, and teaches that the compounds are xanthineoxidase inhibitors and are useful for treatment of gout.

U.S. Pat. No. 4,892,576 discloses pyrazolotriazines of the formula

where X is O or S, Ar is a phenyl, naphthyl, pyridyl or thienyl group,R₆—R₈ are H, alkyl, etc., and R9 is H, alkyl, phenyl, etc. The patentstates that the compounds are useful as herbicides and plant growthregulants.

U.S. Pat. No. 5,484,760 and WO 92/10098 discloses herbicidalcompositions containing, among other things, a herbicidal compound ofthe formula

where A can be N, B can be CR₃, R₃ can be phenyl or substituted phenyl,etc., R is —N(R₄)SO₂R₅ or —SO₂N(R₆)R₇ and R₁ and R₂ can be takentogether to form

where X, Y and Z are H, alkyl, acyl, etc. and D is O or S.

U.S. Pat. No. 3,910,907 and Senga et al., J. Med. Chem., 1982, 25,243-249, disclose triazolotriazines CAMP phosphodiesterase inhibitors ofthe formula

where Z is H, OH, CH₃, C₂H₅, C₆H₅, n-C₃H₇, iso-C₃H₇, SH, SCH₃,NH(n-C-₄H₉), or N(C₂H₅)₂, R is H or CH₃, and R₁ is CH₃ or C₂H₅. Thereference lists eight therapeutic areas where inhibitors of cAMPphosphodiesterase could have utility: asthma, diabetes mellitus, femalefertility control, male infertility, psoriasis, thrombosis, anxiety, andhypertension.

WO95/35298 (Otsuka, 1995) discloses pyrazolopyrimidines and states thatthey are useful as analgesics. The compounds are represented by theformula

where Q is carbonyl or sulfonyl, n is 0 or 1, A is a single bond,alkylene or alkenylene, R¹ is H, alkyl, etc., R² is naphthyl,cycloalkyl, heteroaryl, substituted phenyl or phenoxy, R³ is H, alkyl orphenyl, R⁴ is H, alkyl, alkoxycarbonyl, phenylalkyl, optionallyphenylthio-substituted phenyl, or halogen, R⁵ and R⁶ are H or alkyl.

EP 0 591 528 (Otsuka,1991) discloses anti-inflammatory use ofpyrazolopyrimidines represented by the formula

where R₁, R₂, R₃ and R₄ are H, carboxyl, alkoxycarbonyl, optionallysubstituted alkyl, cycloalkyl, or phenyl, R₅ is SR₆ or NR₇R₈, R₆ ispyridyl or optionally substituted phenyl, and R₇ and R₈ are H oroptionally substituted phenyl.

Springer et al, J. Med. Chem., 1976, vol. 19, no. 2, 291-296 andSpringer U.S. Pat. Nos. 4021,556 and 3,920,652 disclosepyrazolopyrimidines of the formula

where R can be phenyl, substituted phenyl or pyridyl, and their use totreat gout, based on their ability to inhibit xanthine oxidase.

Joshi et al., J. Prakt. Chemie, 321, 2, 1979, 341-344, disclosescompounds of the formula

where R¹ is CF₃, C₂F₅, or C₆H₄F, and R² is CH₃, C₂H₅, CF₃, or C₆H₄F.

Maquestiau et al., Bull. Soc. Belg., vol.101, no. 2, 1992, pages 131-136discloses a pyrazolo[1,5-a]pyrimidine of the formula

Ibrahim et al., Arch. Pharm. (weinheim) 320, 487-491 (1987) disclosespyrazolo[1,5-a]pyrimidines of the formula

where R is NH2 or OH and Ar is 4-phenyl-3-cyano-2-aminopyrid-2-yl.

Other references which disclose azolopyrimidines inclued EP 0 511 528(Otsuka, 1992), U.S. Pat. No. 4,997,940 (Dow, 1991), EP 0 374 448(Nissan, 1990), U.S. Pat. No. 4,621,556 (ICN,1997), EP 0 531 901(Fujisawa, 1993), U.S. Pat. No. 4,567,263 (BASF, 1986), EP 0 662 477(Isagro, 1995), DE 4 243 279 (Bayer, 1994), U.S. Pat. No. 5,397,774(Upjohn, 1995), EP 0 521 622 (Upjohn, 1993), WO 94/109017 (Upjohn,1994), J. Med. Chem., 24, 610-613 (1981), and J. Het. Chem., 22, 601(1985).

SUMMARY OF THE INVENTION

In accordance with one aspect, the present invention provides novelcompounds, pharmaceutical compositions and methods which may be used inthe treatment of affective disorder, anxiety, depression, irritablebowel syndrome, post-traumatic stress disorder, supranuclear palsy,immune suppression, Alzheimer's disease, gastrointestinal disease,anorexia nervosa or other feeding disorder, drug or alcohol withdrawalsymptoms, drug addiction, inflammatory disorder, fertility problems,disorders, the treatment of which can be effected or facilitated byantagonizing CRF, including but not limited to disorders induced orfacilitated by CRF, or a disorder selected from inflammatory disorderssuch as rheumatoid arthritis and osteoarthritis, pain, asthma, psoriasisand allergies; generalized anxiety disorder; panic, phobias,obsessive-compulsive disorder; post-traumatic stress disorder; sleepdisorders induced by stress; pain perception such as fibromyalgia; mooddisorders such as depression, including major depression, single episodedepression, recurrent depression, child abuse induced depression, andpostpartum depression; dysthemia; bipolar disorders; cyclothymia;fatigue syndrome; stress-induced headache; cancer, humanimmunodeficiency virus (HIV) infections; neurodegenerative diseases suchas Alzheimer's disease, Parkinson's disease and Huntington's disease;gastrointestinal diseases such as ulcers, irritable bowel syndrome,Crohn's disease, spastic colon, diarrhea, and post operative ilius andcolonic hypersensitivity associated by psychopathological disturbancesor stress; eating disorders such as anorexia and bulimia nervosa;hemorrhagic stress; stress-induced psychotic episodes; euthyroid sicksyndrome; syndrome of inappropriate antidiarrhetic hormone (ADH);obesity; infertility; head traumas; spinal cord trauma; ischemicneuronal damage (e.g., cerebral ischemia such as cerebral hippocampalischemia); excitotoxic neuronal damage; epilepsy; cardiovascular andhear related disorders including hypertension, tachycardia andcongestive heart failure; stroke; immune dysfunctions including stressinduced immune dysfunctions (e.g., stress induced fevers, porcine stresssyndrome, bovine shipping fever, equine paroxysmal fibrillation, anddysfunctions induced by confinement in chickens, sheering stress insheep or human-animal interaction related stress in dogs); muscularspasms; urinary incontinence; senile dementia of the Alzheimer's type;multiinfarct dementia; amyotrophic lateral sclerosis; chemicaldependencies and addictions (e.g., dependencies on alcohol, cocaine,heroin, benzodiazepines, or other drugs); drug and alcohol withdrawalsymptoms; osteoporosis; psychosocial dwarfism and hypoglycemia in amammal.

The present invention provides novel compounds which bind tocorticotropin releasing factor receptors, thereby altering theanxiogenic effects of CRF secretion. The compounds of the presentinvention are useful for the treatment of psychiatric disorders andneurological diseases, anxiety-related disorders, post-traumatic stressdisorder, supranuclear palsy and feeding disorders as well as treatmentof immunological, cardiovascular or heart-related diseases and colonichypersensitivity associated with psychopathological disturbance andstress in a mammal.

According to another aspect, the present invention provides novelcompounds of Formula (described below) which are useful as antagonistsof the corticotropin releasing factor. The compounds of the presentinvention exhibit activity as corticotropin releasing factor antagonistsand appear to suppress CRF hypersecretion. The present invention alsoincludes pharmaceutical compositions containing such compounds ofFormula (1) and methods of using such compounds for the suppression ofCRF hypersecretion, and/or for the treatment of anxiogenic disorders.

According to yet another aspect of the invention, the compounds providedby this invention (and especially labelled compounds of this invention)are also useful as standards and reagents in determining the ability ofa potential pharmaceutical to bind to the CRF receptor.

DETAILED DESCRIPTION OF INVENTION

[1] In a first embodiment the present invention provides for a compoundof Formula (I) or (II)

wherein:

D is Ar¹ or heteroaryl, each optionally substituted with 1 to 5 R⁵groups and each is attached to an unsaturated carbon atom;

R¹ is H, Ar², heteroaryl, heterocyclyl, or carbocyclyl; or

C₁-C₆ alkyl, C₂-C₄ alkenyl, or C₂-C₄ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₄ alkyl, Ar², heteroaryl, heterocyclyl, carbocyclyl,OR¹², F, Cl, Br, I, CF₃, and NO₂;

R² is H, Ar², heteroaryl, heterocyclyl, or carbocyclyl; or

C₁-C₁₀ alkyl, C₃-C₁₀ alkenyl, C₃-C₁₀ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, Ar²,heteroaryl, heterocyclyl, carbocyclyl, F, Br, Cl, I, CN, OR¹², SR¹⁵,NO₂, NR⁹COR¹³, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰, and CONR¹¹R¹⁰;

R³ is H, Ar², heteroaryl, heterocyclyl, or carbocyclyl; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, Ar², heteroaryl, heterocyclyl,carbocyclyl, NO₂, F, Cl, Br, I, NR⁹COR⁷, NR⁹CO₂R⁷, OR⁷, CONR¹⁰R¹¹, andCO(NOR¹²)R⁷;

R⁴ is NR₁₁R¹⁰;

R⁵ is independently selected at each occurrence from:

NO₂, F, Cl, Br, I, CN, NR¹⁰R¹¹, NR⁹COR¹³, NR⁹CO₂R⁷, COR¹³, R¹³, OR¹²,CONR¹⁰R¹¹, CO(NOR⁹)R¹⁰, CO₂R¹², and S(O)_(n)R¹⁴; or

C₁-C₆ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₄ alkyl, C₃-C₆ cycloalkyl, CF₃, NO₂, F, Cl, Br, I,CN, NR⁶R⁷, NR⁹COR⁷, NR⁹CO₂R⁷, COR⁷, OR⁷, CONR⁶R⁷, CO₂R⁷, CO(NOR⁹)R⁷, andS(O)_(n)R⁷;

R⁶ is independently selected at each occurrence from:

H, —CH₂Ar², Ar², heteroaryl, heterocyclyl, and carbocyclyl; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,OR¹², NO₂, S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹⁴, NR⁹COR¹³, N(COR¹³)₂,NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰, CONR¹¹R¹⁰, Ar², heteroaryl,heterocyclyl, and carbocyclyl;

R⁷ is independently selected at each occurrence from:

H, —CH₂Ar², Ar², heteroaryl, heterocyclyl, and carbocyclyl; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,OR¹², NO₂, S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂,NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰, CONR¹¹R¹⁰, Ar², heteroaryl,heterocyclyl, and carbocyclyl;

Ar¹ is phenyl or naphthyl;

Ar² is phenyl or naphthyl, each optionally substituted with 1 to 5substituents independently selected at each occurrence from C₁-C₆ alkyl,C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I, OR¹², NO₂,S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰,NR⁹CO₂R¹², NR¹¹R¹⁰, and CONR¹¹R¹⁰;

heteroaryl is pyridyl, pyrimidinyl, triazinyl, furanyl, pyrrolyl,imidazolyl, pyranyl, quinolinyl, isoquinolinyl, thienyl, imidazolyl,thiazolyl, indolyl, indazolyl, pyrrolyl, oxazolyl, benzofuranyl,benzothienyl, benzothiazolyl, isoxazolyl, pyrazolyl,2,3-dihydrobenzothienyl or 2,3-dihydrobenzofuranyl;

heterocyclyl is saturated or partially saturated heteroaryl, optionallysubstituted with 1 to 5 substituents independently selected at eachoccurrence from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, CN, F,Cl, Br, I, OR¹², NO₂, S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³, NR⁹COR¹³,N(COR¹³)₂, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹⁰R¹¹, and CONR¹¹R¹⁰;

carbocyclyl is saturated or partially unsaturated C₃-C₁₀ membered ring,optionally substituted with 1 to 5 substituents independently selectedat each occurrence from C₁-C₆ alkyl, CF₃, CN, F, Cl, Br, I, OR¹², SR¹⁰,S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰,NR⁹CO₂R¹², NR¹⁰R¹¹, and CONR¹¹R¹⁰;

R⁹ is independently selected at each occurrence from H, C₁-C₄ alkyl, andC₃-C₆ cycloalkyl;

R¹⁰ is H, heterocyclyl, or carbocycle; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, Ar², heteroaryl, eachoptionally substituted with 1-3 F, Cl, Br, I, NO₂, CF3, CN, or OR¹²;

R¹¹ is H, heterocyclyl, or carbocycle; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, and C₂-C₆ alkynyl, Ar², heteroaryl, eachoptionally substituted with 1-3 C₁-C₆ alkyl, C₃-C₆ cycloalkyl, F, Cl,Br, I, NO₂, CF3, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 C₁-C₄ alkyl groups;

R¹² is independently selected at each occurrence from H, C₁-C₆ alkyl,C₃-C₆ cycloalkyl, and C₁-C₄ haloalkyl;

R¹³ is selected from H, C₁-C₄ alkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy C₁-C₆alkyl, C₃-C₆ cycloalkyl, and C₃-C₆ cycloalkyl C₁-C₆ alkyl;

R¹⁴ is independently selected at each occurrence from C₁-C₆ alkyl, C₃-C₆alkyl, and phenyl, each subsituted by 1-3 C₁-C₄ alkyl, C₁-C₄ haloalkylor OR¹⁵; and

R¹⁵ is independently selected at each occurrence from H, C₁-C₆ alkyl,C₃-C₆ cycloalkyl, and C₁-C₄ haloalkyl.

[2] In a more preferred embodiment, the present invention provides for acompound of Formula (Ia),

wherein

A is CR¹³ or N;

B is CR¹³ or N;

n is 0, 1, 2, or 3;

R¹ is H; or

C₁-C₆ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, each optionally substitutedwith 1 to 3 substituents independently selected at each occurrence fromC₁-C₄ alkyl, OR¹², F, Cl, Br, I, CF₃, and NO₂;

R² is H; or

C₁-C₁₀ alkyl, C₃-C₁₀ alkenyl, C₃-C₁₀ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, F, Br,Cl, I, CN, OR¹², SR¹⁵, NO₂, NR⁹COR¹³, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰,and CONR¹¹R¹⁰;

R₃ is H; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, or C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, NO₂, F, Cl, Br, I, NR⁹COR⁷,NR⁹CO₂R⁷, OR⁷, CONR¹⁰R¹¹, or CO(NOR¹²)R⁷;

R⁴ is NR¹⁰R¹¹;

R⁶ is independently selected at each occurrence from:

H, —CH₂Ar², and Ar²; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, and C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,NO₂, and OR¹²;

R⁷ is independently selected at each occurrence from:

H, —CH₂Ar², and Ar²; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,NO₂, and OR¹²;

Ar² is phenyl optionally substituted with 1 to 5 substituentsindependently selected at each occurrence from C₁-C₆ alkyl, C₃-C₆cycloalkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I, OR¹², and NO₂;

R¹⁰ is H; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, Ar², heteroaryl, eachoptionally substituted with 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

R¹¹ is H; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, and C₂-C₆ alkynyl, Ar², heteroaryl, eachoptionally substituted with 1-3 C₁-C₆ alkyl, C₃-C₆ cycloalkyl, F, Cl,Br, I, NO₂, CF₃, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 C₁-C₄ alkyl groups.

[3] In a further more preferred embodiment, the present inventionprovides for a compound of Formula (Ia),

wherein

A is CR¹³;

B is CR¹³;

n is 0, 1, 2, or 3;

R¹ is H; or

C₁-C₄ alkyl, C₂-C₄ alkenyl, or C₂-C₄ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₄ alkyl, OR¹², F, Cl, Br, I, CF₃, and NO₂;

R² is H; or

C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, each optionally substitutedwith 1 to 3 substituents independently selected at each occurrence fromC₁-C₄ alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, F, Br, Cl, I, CN, OR¹²,SR¹⁵, NO₂, NR⁹CO₂R¹², NR¹¹R¹⁰, and CONR¹¹R¹⁰;

R³ is H; or

C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, NO₂, F, Cl, Br, I, NR⁹COR⁷,NR⁹CO₂R⁷, OR⁷, CONR¹⁰R¹¹, and CO(NOR¹²)R⁷;

R⁴ is NR¹⁰R¹¹;

R¹⁰ is H; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, each optionally substitutedwith 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

R¹¹ is H; or

C₁-C₆ alkyl, C₂-C₆ alkenyl, and C₂-C₆ alkynyl, each optionallysubstituted with 1-3 C₁-C₆ alkyl, C₃-C₆ cycloalkyl, F, Cl, Br, I, NO₂,CF₃, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 C₁-C₄ alkyl groups.

[4] In an even more preferred embodiment, the present invention providesfor a compound of Formula (Ib)

wherein

n is 1, 2, or 3;

R⁵ is independently selected at each occurrence from:

NO₂, F, Cl, Br, I, CN, NR⁹COR¹³, NR⁹CO₂R⁷, COR¹³, R¹³, CONR¹OR¹¹,CO(NOR⁹)R¹⁰, CO₂R¹², and S(O)_(n)R¹⁴;

R⁴ is NR¹⁰R¹¹;

R¹⁰ is H; or

C₁-C₄ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, each optionally substitutedwith 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

R¹¹ is H; or

C₁-C₄ alkyl, C₂-C₄ alkenyl, and C₂-C₄ alkynyl, each optionallysubstituted with 1-3 C₁-C₄ alkyl, C₃-C₆ cycloalkyl, F, Cl, Br, I, NO₂,CF₃, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-2 C₁-C₄ alkyl groups.

[5] In another preferred embodiment, the present invention provides fora compound of Formula (Ic),

wherein

R⁵ is independently selected at each occurrence from: NO₂, F, Cl, Br, I,CN, and R¹³;

R⁴ is NR¹⁰R¹¹;

R¹⁰ is H; or

methyl, ethyl, propyl, butyl, ethene, propene, butene, propargyl, eachoptionally substituted with 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

R¹¹ is H; or

methyl, ethyl, propyl, butyl, ethene, propene, each optionallysubstituted with 1-2 methyl, ethyl, propyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 methyl, ethyl, or propyl groups.

[6] In another more preferred embodiment, the present invention providesfor a compound of Formula (IIa),

wherein:

R¹ is H; or

C₁-C₆ alkyl, C₂-C₄ alkenyl, C₂-C₄ alkynyl, each optionally substitutedwith 1 to 3 substituents independently selected at each occurrence fromC₁-C₄ alkyl, OR¹², F, Cl, Br, I, CF₃, and NO₂;

R³ is H; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, or C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, NO₂, F, Cl, Br, I, NR⁹COR⁷,NR⁹CO₂R⁷, OR⁷, CONR¹OR¹¹, or CO(NOR¹²)R⁷;

R⁴ is NR¹¹R¹⁰;

R⁵ is independently selected at each occurrence from:

NO₂, F, Cl, Br, I, CN, and R¹³;

R⁷ is independently selected at each occurrence from:

H, —CH₂Ar², and Ar²; or

C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,NO₂, and OR¹²;

Ar² is phenyl optionally substituted with 1 to 5 substituentsindependently selected at each occurrence from C₁-C₆ alkyl, C₃-C₆cycloalkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I, OR¹², and NO₂;

R⁹ is independently selected at each occurrence from H, C₁-C₄ alkyl, andC₃-C₆ cycloalkyl;

R¹⁰ is independently selected at each occurrence from H, C₁-C₆ alkyl,C₂-C₆ alkenyl, and C₂-C₆ alkynyl, each optionally substituted with 1-3F, Cl, Br, I, NO₂, CF₃, or OR¹²;

R¹¹ is independently selected at each occurrence from H, C₁-C₆ alkyl,C₂-C₆ alkenyl, and C₂-C₆ alkynyl, each optionally substituted with 1-3C₁-C₆ alkyl, C₃-C₆ cycloalkyl, F, Cl, Br, I, NO₂, CF₃, or OR¹²;alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 C₁-C₄ alkyl groups;

R¹² is independently selected at each occurrence from H, C₁-C₆ alkyl,C₃-C₆ cycloalkyl, and C₁-C₄ haloalkyl;

R¹³ is selected from H, C₁-C₄ alkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy C₁-C₆alkyl, C₃-C₆ cycloalkyl, and C₃-C₆ cycloalkyl C₁-C₆ alkyl.

[7] In another further more preferred embodiment, the present inventionprovides for a compound of Formula (IIa), wherein:

R⁵ is independently selected at each occurrence from:

NO₂, F, Cl, Br, I, CN, and R¹³;

R⁴ is NR¹⁰R¹¹;

R¹⁰ is H; or

methyl, ethyl, propyl, butyl, ethene, propene, butene, propargyl, eachoptionally substituted with 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

R¹¹ is H; or

methyl, ethyl, propyl, butyl, ethene, propene, each optionallysubstituted with 1-2 methyl, ethyl, propyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;

alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 methyl, ethyl, or propyl groups.

[8] In another even further more preferred embodiment, the compounds ofthe present invention are selected from:

4-[bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5,6-trimethylpyrrolo[3,4-d]pyrimidine;

4-[bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine;

4-(N,N-diethylamino)-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidine;

4-(N-cyclopropylmethyl-N-propylamino)-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidine;

4-(N-butyl-N-ethylamino)-7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine;

4-[bis(cyclopropylmethyl)amino]7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine; and

7-(2,4-Dichloro-phenyl)-4-(1-ethyl-propylamino)-2,5-dimethyl-5H-pyrrolo[3,4-d]pyrimidin-5-ol.

[9] In a second embodiment the present invention provides for apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound of formula(Ia) or (IIa).

[10] In a third embodiment the present invention provides for a methodof treating affective disorder, anxiety, depression in a mammalcomprising administering to the mammal a dose of the composition of acompound of Formula (Ia) or (IIa).

Many compounds of this invention have one or more asymmetric centers orplanes. Unless otherwise indicated, all chiral (enantiomeric anddiastereomeric) and racemic forms are included in the present invention.Many geometric isomers of olefins, C═N double bonds, and the like canalso be present in the compounds, and all such stable isomers arecontemplated in the present invention. The compounds may be isolated inoptically active or racemic forms. It is well known in the art how toprepare optically active forms, such as by resolution of racemic formsor by synthesis from optically active starting materials. All chiral,(enantiomeric and diastereomeric) and racemic forms and all geometricisomeric forms of a structure are intended, unless the specificstereochemistry or isomer form is specifically indicated.

The term “alkyl” includes both branched and straight-chain alkyl havingthe specified number of carbon atoms. Commonly used abbreviations havethe following meanings: Me is methyl, Et is ethyl, Pr is propyl, Bu isbutyl. The prefix “n” means a straight chain alkyl. The prefix “c” meansa cycloalkyl. The prefix “(S)” means the S enantiomer and the prefix“(R)” means the R enantiomer. Alkenyl” includes hydrocarbon chains ofeither a straight or branched configuration and one or more unsaturatedcarbon—carbon bonds which may occur in any stable point along the chain,such as ethenyl, propenyl, and the like. “Alkynyl” includes hydrocarbonchains of either a straight or branched configuration and one or moretriple carbon—carbon bonds which may occur in any stable point along thechain, such as ethynyl, propynyl and the like. “Haloalkyl” is intendedto include both branched and straight-chain alkyl having the specifiednumber of carbon atoms, substituted with 1 or more halogen; “alkoxy”represents an alkyl group of indicated number of carbon atoms attachedthrough an oxygen bridge; “cycloalkyl” is intended to include saturatedring groups, including mono-, bi- or poly-cyclic ring systems, such ascyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and so forth. “Halo”or “halogen” includes fluoro, chloro, bromo, and iodo.

The term “substituted”, as used herein, means that one or more hydrogenon the designated atom is replaced with a selection from the indicatedgroup, provided that the designated atom's normal valency is notexceeded, and that the substitution results in a stable compound. When asubstitent is keto (i.e., ═O), then 2 hydrogens on the atom arereplaced.

Combinations of substituents and/or variables are permissible only ifsuch combinations result in stable compounds. By “stable compound” or“stable structure” is meant a compound that is sufficiently robust tosurvive isolation to a useful degree of purity from a reaction mixture,and formulation into an efficacious therapeutic agent.

The term “appropriate amino acid protecting group” means any group knownin the art of organic synthesis for the protection of amine orcarboxylic acid groups. Such amine protecting groups include thoselisted in Greene and Wuts, “Protective Groups in Organic Synthesis” JohnWiley & Sons, New York (1991) and “The Peptides: Analysis, Synthesis,Biology, Vol. 3, Academic Press, New York (1981), the disclosure ofwhich is hereby incorporated by reference. Any amine protecting groupknown in the art can be used. Examples of amine protecting groupsinclude, but are not limited to, the following: 1) acyl types such asformyl, trifluoroacetyl, phthalyl, and p-toluenesulfonyl; 2) aromaticcarbamate types such as benzyloxycarbonyl (Cbz) and substitutedbenzyloxy-carbonyls, 1-(p-biphenyl)-1-methylethoxy-carbonyl, and9-fluorenylmethyloxycarbonyl (Fmoc); 3) aliphatic carbamate types suchas tert-butyloxycarbonyl (Boc), ethoxycarbonyl,diisopropylmethoxycarbonyl, and allyloxycarbonyl; 4) cyclic alkylcarbamate types such as cyclopentyloxycarbonyl and adamantyloxycarbonyl;5) alkyl types such as triphenylmethyl and benzyl; 6) trialkylsilanesuch as trimethylsilane; and 7) thiol containing types such asphenylthiocarbonyl and dithiasuccinoyl.

The term “pharmaceutically acceptable salts” includes acid or base saltsof the compounds of Formulae (1) and (2). Examples of pharmaceuticallyacceptable salts include, but are not limited to, mineral or organicacid salts of basic residues such as amines; alkali or organic salts ofacidic residues such as carboxylic acids; and the like.

Pharmaceutically acceptable salts of the compounds of the invention canbe prepared by reacting the free acid or base forms of these compoundswith a stoichiometric amount of the appropriate base or acid in water orin an organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. Lists of suitable salts are found in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,1985, p. 1418, the disclosure of which is hereby incorporated byreference.

“Prodrugs” are considered to be any covalently bonded carriers whichrelease the active parent drug of formula (I) in vivo when such prodrugis administered to a mammalian subject. Prodrugs of the compounds offormula (I) are prepared by modifying functional groups present in thecompounds in such a way that the modifications are cleaved, either inroutine manipulation or in vivo, to the parent compounds. Prodrugsinclude compounds wherein hydroxy, amine, or sulfhydryl groups arebonded to any group that, when administered to a mammalian subject,cleaves to form a free hydroxyl, amino, or sulfhydryl group,respectively. Examples of prodrugs include, but are not limited to,acetate, formate and benzoate derivatives of alcohol and aminefunctional groups in the compounds of formula (I); and the like.

The term “therapeutically effective amount” of a compound of thisinvention means an amount effective to antagonize abnormal level of CRFor treat the symptoms of affective disorder, anxiety or depression in ahost.

Syntheses

A pyrrolo[3,4-d]pyrimidine of Formula (I′) may be prepared from anintermediate X using the procedures outlined in Scheme 1. A compound Xmay be treated with a halogenating agent in the presence or absence of abase in the presence or absence of an inert solvent at reactiontemperatures ranging from −80° C. to 250° C. to give a product XI (whereL is halogen). Halogenating agents include, but are not limited to,Cu(II)L₂ (L=halogen), Br₂, Cl₂, 12, N-bromosuccinimide,N-iodosuccinimide or N-chlorosuccinimide. Bases may include, but are notlimited to, alkali metal carbonates, alkali metal bicarbonates, trialkylamines (preferably N,N-di-isopropyl-N-ethyl amine) or aromatic amines(preferably pyridine). Inert solvents may include, but are not limitedto, lower alkanenitriles (1 to 6 carbons, preferably acetonitrile),dialkyl ethers (preferably diethyl ether), cyclic ethers (preferablytetrahydrofuran or 1,4-dioxane), N,N-dialkylformamides (preferablydimethylformamide), N,N-dialkylacetamides (preferablydimethylacetamide), cyclic amides (preferably N-methylpyrrolidin-2-one),dialkylsulfoxides (preferably dimethylsulfoxide), aromatic hydrocarbons(preferably benzene or toluene), alkyl esters (preferably EtOAc) orhaloalkanes of i to 10 carbons and 1 to 10 halogens (preferablydichloromethane). Preferred reaction temperatures range from −20° C. to150° C. The resulting intermediate XI may then be reacted with beta ketoester derivative XII in the presence of a base such as alkali metalalkoxides in a solvent such as aliphatic alcohols or an inert solvent attemperatures ranging from −20° C. to 150° C. to give a product XIII.Inert solvents may include, but are not limited to, polyethers(preferably 1,2-dimethoxyethane), dialkyl ethers (preferably diethylether), cyclic ethers (preferably tetrahydro-furan or 1,4-dioxane) oraromatic hydrocarbons (preferably benzene or toluene). Intermediate XIIIis treated with a ammonium salts (preferably ammonium acetate) in anorganic acid medium (preferably acetic acid) at temperatures rangingfrom −20° C. to 150° C. to provide a compound XIV. The pyrrole nitrogenof compound XIV may be alkylated using an R²LG group in presence of basein an inert solvent to afford a compound XV. LG is a leaving group(which may be a halide, tosylate or a mesylate. Bases may include, butare not limited to, alkali metal hydrides (preferably sodium hydride).

Inert solvents include, but are not limited to, dialkyl ethers(preferably diethyl ether), cyclic ethers (preferably tetrahydrofuran or1,4-dioxane), N,N-dialkylformamides (preferably dimethylformamide),N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides (preferablydimethylsulfoxide) or aromatic hydrocarbons (preferably benzene ortoluene). Preferred reaction temperatures range from −20° C. to 100° C.The resulting pyrrole derivative XV may be converted to aza derivativeXVI by treatment with an aryl diazonium salts in the presence of analkali metal salts (preferably NaOAc but not limited to) in an acidmedium (preferably AcOH but not limited to). The compound of Formula XVImay be reduced to its amino derivative XVII using tin (II) halides (butnot limited to) in an acid medium. Acids may include, but are notlimited to alkanoic acids of 2 to 10 carbons (preferably acetic acid),haloalkanoic acids (2-10 carbons, 1-10 halogens, such as trifluoroaceticacid), alkanesulfonic acids of 1 to 10 carbons (preferablymethanesulfonic acid), hydrochloric acid. Compound XVII may be convertedto amidine derivative XVIII by treatment with a nitrile R¹CN in thepresence of anhydrous acids (preferably HCl gas but not limited to) inan inert solvent at ambient temperature. Inert solvents may include, butare not limited to, cyclic ethers (preferably tetrahydrofuran or1,4-dioxane) or aromatic hydrocarbons (preferably benzene or toluene).The amidine derivative XVIII may be cyclized to pyrrolopyrimidonederivative XIX by treatment with either anhydrous or aqeous ammonia insolvents such as aliphatic alcohol. Compound XX may be obtained bytreament of compound XIX with halogenating agents in the presence orabsence of a base and inert solvents. Halogenating agents include, butare not limited to, POCl₃, SOCl₂, PCl₃, PCl₅ or PBr₃. Inert solvents forthe halogenation include, but are not limited to, aromatic hydrocarbons(preferably benzene or toluene), or haloalkanes of 1 to 10 carbons and 1to 10 halogens (preferably dichloromethane). Preferred reactiontemperatures range from 0° C. to 150° C. Bases may include, but are notlimited to, alkali metal carbonates, alkali metal bicarbonates, trialkylamines (preferably N,N-di-isopropyl-N-ethyl amine) or aromatic amines(preferably N,N-alkylamines). Compound of the Formula XX may beconverted to a compound of present invention (I′) by treatment with anamine HNR¹¹R¹² in the presence or absence of a base as well as in thepresence or absence of an inert solvent at temperatures ranging from−80° C. to 150° C.

Alternatively, compounds of the Formula (I′) may be obtained fromintermediate XXI as described in Scheme 2. An aryl alpha aminonitrileXXII may be prepared by reacting aromatic aldehydes with cyanidereagents, but not limited to trialkylsilylcyanide in the presence of alewis acids such as zinc iodide (but not limited to) in

an inert solvent such as haloalkanes (but not limited to). The resultingsilylether derivative may be converted to compounds of Formula XXII bytreatment with anhydrous ammonia in aliphatic alcohols (but not limitedto). The resulting intermediates XXII may be converted to a compoundXXIII by treatment with a beta keto ester derivative XII in the presenceof an acid in an inert solvent at temperatures ranging from −20° C. to150° C. Inert solvents may include, but are not limited to, polyethers(preferably 1,2-dimethoxyethane), dialkyl ethers (preferably diethylether), cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane) oraromatic hydrocarbons (preferably benzene or toluene). Acids mayinclude, but are not limited to alkanoic acids of 2 to 10 carbons(preferably acetic acid), haloalkanoic acids (2-10 carbons, 1-10halogens, such as trifluoroacetic acid), arylsulfonic acids (preferablyp-toluenesulfonic acid or benzenesulfonic acid), alkanesulfonic acids of1 to 10 carbons (preferably methanesulfonic acid) or hydrochloric acid.The pyrrole derivative XXIV may be obtained by treatment of compoundXXIII with a base in an inert solvent. Bases may include, but are notlimited to, alkali metal hydrides (preferably sodium hydride), alkalimetal alkoxides, alkali metal dialkylamides (preferably lithiumdi-isopropylamide) and alkali metal bis(trialkylsilyl)-amides(preferably sodiumbis(trimethylsilyl) amide). Inert solvents include,but are not limited to, dialkyl ethers (preferably diethyl ether),cyclic ethers (preferably tetrahydrofuran or 1,4-dioxane), aliphaticalcohols, N,N-dialkylformamides (preferably dimethylformamide),N,N-dialkylacetamides (preferably dimethylacetamide), cyclic amides(preferably N-methylpyrrolidin-2-one), dialkylsulfoxides (preferablydimethylsulfoxide) or aromatic hydrocarbons (preferably benzene ortoluene). Compounds XXIV may then be converted to Formula (I′) byfollowing similar conditions outlined in Scheme 1.

EXAMPLES

Analytical data were recorded for the compounds described below usingthe following general procedures. Proton NMR spectra were recorded on anVarian FT-NMR (300 MHz); chemical shifts were recorded in ppm (δ) froman internal tetramethysilane standard in deuterochloroform ordeuterodimethylsulfoxide as specified below. Mass spectra (MS) or highresolution mass spectra (HRMS) were recorded on a Finnegan MAT 8230spectrometer (using chemical ionization (CI) with NH₃ as the carrier gasor gas chromatography (GC) as specified below) or a Hewlett Packard5988A model spectrometer. Melting points were recorded on a Buchi Model510 melting point apparatus and are uncorrected. Boiling points areuncorrected. All pH determinations during workup were made withindicator paper.

Reagents were purchased from commercial sources and, where necessary,purified prior to use according to the general procedures outlined by D.Perrin and W. L. F. Armarego, Purification of Laboratory Chemicals, 3rded., (N.Y.: Pergamon Press, 1988). Chromatography (thin layer (TLC) orpreparative) was performed on silica gel using the solvent systemsindicated below. For mixed solvent systems, the volume ratios are given.Otherwise, parts and percentages are by weight.

The following examples are provided to describe the invention in furtherdetail. These examples, which set forth the best mode presentlycontemplated for carrying out the invention, are intended to illustrateand not to limit the invention.

Example 14-[bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5,6-trimethylpyrrolo[3,4-d]pyrimidine

Part A: To an ice cold solution of ethyl acetoacetate (13.0 g, 0.1 mole)in tetrahydrofuran (200 mL) was added dropwise 1.0 M NaHMDS in THF (100mL, 0.1 mole) over 30 mins under nitrogen atmosphere. After the additionthe reaction mixture further stirred at 0° C. for additional 15 mins. Tothis cold stirred solution was added dropwise 2-bromo-2′,4‘-’dichloroacetophenone (26.0 g, 0.1 mole, prepared according to D. M.Rotstein, D. J. Kertesz, K. A. M. Walker, D.C. Swinney, J.Med. Chem.1992, 35, 2818-2825) in THF (50 mL) over 15 mins. Stirring was continuedfor additional 2 h at 0° C. and TLC (dichloromethane) revealed absenceof starting material spot (Rf=0.88) and a new spot was noticed(Rf=0.56). The reaction mixture was quenched with water (10 mL) and thesolvent was evaporated in vacuo. The residue was diluted with water (500mL), extracted with EtOAc (3×250 mL), washed the combined organic layerswith brine (300 mL) and dried with anhydrous magnesium sulfate. Thedried extract was filtered and concentrated in vacuum to afford aresidue. The residue was purified by flash column chromatography on asilica gel using dichloromethane as a eluent to afford the dicarbonylester derivative (21.3 g, 67% yield) as a pale yellow oil. Anal. calcd.for C₁₄H₁₄O₄Cl₂: C, 53.02; H, 4.46. Found: C, 52.95; H, 4.27.

Part B: Ethyl 2-(2,4-dichlorophenyl)-5-methyl-pyrrole carboxylate: Thecompound of part A of Example 1 (10.8 g, 0.034 moles) in glacial aceticacid (250 mL) was treated with anhydrous ammonium acetate (26.2 g, 0.34moles, 10 equiv.) and refluxed under nitrogen atmosphere for 48 h. Thereaction mixture was then cooled to room temperature and poured overcrushed ice (750 g) and stirred for 1 h. The light pink colored solidseparated was filtered, dried and purified by flash columnchromatography on a silica gel using 0.5% MeOH/CH₂Cl₂ as eluent affordeddesired pyrrole derivative (9.4 g, mp 144-145° C., 93% yield). Anal.calcd. for C₁₄H₁₃Cl₂NO₂: C, 56.40; H, 4.39; N, 4.71. Found: C, 56.11; H,4.11; N, 4.54.

Part C: Ethyl 2-(2,4-dichlorophenyl)-1,5-dimethyl-pyrrole carboxylate:The compound of part B of Example 1 (5.96 g, 0.02 moles) was dissolvedin anhydrous DMF (100 mL) and treated with 60% NaH (0.96 g, 0.024 moles)at 0° C. under nitrogen atmosphere. The mixture was stirred for 15 minsand then treated with excess of iodomethane. The mixture was slowlybrought to room temperature and stirred at room temperature for 3 h. TLC(1% MeOH/CH₂Cl₂) revealed absence of starting material spot (Rf=0.56)and a new spot was noticed (Rf=0.75). Later the reaction mixture wasquenched with ice, extracted with EtOAc (3*75 mL), washed the combinedorganic layers with brine (50 mL) and dried with anhydrous magnesiumsulfate. The dried extract was filtered and concentrated in vacuum toafford a residue. The residue was purified by flash columnchromatography on a silica gel using dichloromethane as eluent affordedN-methylpyrrole derivative (6.0 g, mp 69-70° C., 96% yield) as a whitesolid. Anal. calcd. for C₁₅H₁₅Cl₂NO₂: C, 57.71; H, 4.849; N, 4.50.Found: C, 57.53; H, 4.71; N, 4.35.

Part D: Ethyl 3-amino-2-(2,4-dichlorophenyl)-1,5-dimethylpyrrolecarboxylate: To a solution of 4-nitroaniline (2.76 g, 0.02 moles) in 6 Naqeous HCl(15 mL) at −5° C. was added dropwise a solution of NaNO₂ (2.25g, 0.0325 moles) in water (7.5 mL) over 15 mins. After the addition thecompound of part C of Example 1 (5.0 g, 0.016 moles) was in dissolved inAcOH (80 mL) and NaOAc (4.0 g)and added to the reaction mixture at 0° C.over 60 mins. Stirring was continued for additional 30 mins at 0° C. andthen poured over crushed ice (200 g). The solid separated was filtered,washed with water, dried and purified by flash column chromatography toafford 0.91 g of desired 3-diazapyrrole derivative (17% yield, based on1.4 g of recovered starting material pyrrole derivative from the column)as red orange liquid. Tin(II) chloride (3.8 g, 20 mmole) was suspendedin glacial AcOH (4.0 mL) and heated to 80° C. with stirring. To thismixture was added 3-diazapyrrole derivative (0.91 g, 2.0 mmole)dissolved in AcOH (6.0 mL) over a period of 15 mins. at 80° C. Theresultant slurry was stirred for a period of 4 h at 80° C.TLC (CH₂Cl₂)indicated absence of starting material (Rf=0.2) and a new spot wasnoticed (Rf=0.5). The solvent from the reaction mixture was removed andthe residue was poured over crushed ice, adjusted the pH to 8 to 9 usingdilute NaOH, treated the mixture with 50 mL of EtOAc, filtered theresultant turbid solution over celite, washed the celite with 2×25 mL ofEtOAc. The aqeous layer from the filterate was separated from organiclayer, washed with brine (25 mL) and dried (MgSO₄). The solvent wasstripped in vacuo, and the residue was passed through flash column(eluent: 0.5% methanol/CH₂Cl₂) and the oil obtained was crystallizedfrom diethyl ether to afford (280 mg, mp 115-116° C., 43% yield) paleyellow crystalline solid. Anal. calcd. for C₁₅H₁₆Cl₂N₂O₂: C, 55.06; H,4.949; N,8.56. Found: C, 55.09; H, 4.99; N, 8.50.

Part E:7-(2,4-Dichlorophenyl)-2,5,6-trimethyl-pyrrolo[3,4-d]pyrimidine-4(3H)-one:The compound of part D of Example 1 (0.25 g, 0.764 mmol) was dissolvedin dioxane (2.0 mL) and acetonitrile (2.0 mL) and cooled to 15° C. undernitrogen. Dry HCl (gas) was passed through a syringe needle into thereaction mixture over a period of 4 h. TLC(eluent: 10:1 CH₂Cl₂/methanol)revealed presence of two new spots at Rf=0.27 and 0.46 (faint) alongwith unreacted starting material spot at Rf=0.80. Stopped bubbling HClgas at this stage and allowed to stir at room temp. over night. TLC(eluent: 10:1 CH₂Cl₂/methanol) revealed absence of starting materialspot and also the faint spot at Rf=0.46. The only spot noticed was atRf=0.27 and mass spectrum revealed formation of acetonitrile additionproduct (M+H=368). The solvent was removed under vacuum, residue wasdissolved in 4.0 mL of 1:1 EtOH/water and basified using 28% ammoniumhydroxide to pH 9 to 10. Some solid separation was noticed, butextraction of the mixture with ethyl acetate (15 mL) resulted inseparation of white solid. Filtered the solid, washed with ethylacetate, and dried to afford (0.159 g) desired product. Additional 60 mgof product was obtained by extraction of the aq. layer with EtOAc.Overall yield 0.22 g (88% yield, mp>290° C.). Anal. calcd. forC₁₅H₁₃Cl₂N₃O: C, 55.92; H, 4.089; N,13.04. Found: C, 55.77; H, 3.99; N,12.80.

Part F:4-Chloro-7-(2,4-dichlorophenyl)-2,5,6-trimethyl-pyrrolo[3,4-d]pyrimidine:The compound of part E of Example 1 (0.125 g, 0.39 mmol) was treatedwith POCl₃ (2.0 mL) and heated at 80° C. for 48 h. Excess POCl₃ wasremoved under vacuum and then quenched with ice (10 g). The reactionmixture was then extracted extracted with EtOAc (3×15 mL), washed thecombined organic layers with brine (50 mL) and dried with anhydrousmagnesium sulfate. The dried extract was filtered and concentrated invacuum to afford a residue. The residue was purified by flash columnchromatography on a silica gel using 1% MeOH/dichloromethane as eluentto afford corresponding chloro derivative (70 mg, mp 191-192° C., 54%yield) as a yellow solid.

Part G:4-[bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5,6-trimethyl-pyrrolo[3,4-d]pyrimidine:The compound of part F of Example 1 (0.57 mg, 0.168 mmol) in ethanol(2.0 mL) was treated with bis (2-methoxyethyl)amine (67 mg, 0.503 mmol,3 eq.) and heated at 80° C. for 20 h. The reaction mixture wasconcentrated in vacuum to afford a residue. The residue was purified byflash column chromatography on a silica gel using 2%MeOH/dichloromethane as eluent to afford desired product (43.8 mg, 59%yield) as a yellow oil. HRMS calcd. for C₂₁H₂₇Cl₂N₄O₂: 437.1511. Found:437.1497.

Example 24-[Bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine

Part A: •-Amino-2,4-dichlorophenylacetonitrile: To a solution of2,4-dichlorobenzaldehyde (35.0 g, 0.2 moles) in dichloromethane (200.0mL) was added anhydrous zinc iodide (1.0 g) followed by dropwiseaddition of TMSCN (25.0 g, 0.25 moles) at room temperature. Theresultant mixture was stirred for 30 mins and examination by NMRrevealed formation of alpha-silyloxynitrile as a clean product (Productnot UV visible). The solvent was evaporated in vacuum and the residuewas treated 7.0 M ammonia in methanol (200 mL) and continued thereaction at 40° C. for 16 h. TLC (2% MeOH/CH₂Cl₂) indicated a new spot(Rf=0.27, visible only with iodine). The solvent was evaporated and thecrude was purified by flash column chromatography (eluent CH₂Cl₂) toafford desired product as a yellow solid (34.75 g, mp 74-75° C., 87%yield). Anal. calcd. for C₈H₆Cl₂N₂: C, 47.79; H, 3.02; N,13.93. Found:C, 47.94; H, 3.12; N, 13.71.

Part B: Ethyl N-(alpha-cyano-2,4-dichlorobenzyl)-3-amino-2-butenoate: Toa solution of alpha-amino-2,4-dichlorophenylacetonitrile (20.1 g, 0.1mole) in benzene (250 mL) was treated with ethyl acetoacetate (13.0 g,0.1 mole) and p-toluenesulfonic acid monohydrate (0.475 g, 0.0025 mole).The reaction mixture was refluxed for 48 h. TLC (eluent 2% MeOH/CH₂Cl₂)revealed trace amount of starting material nitrile (Rf=0.27; visibleonly under iodine) along with a new spot (Rf=0.8). The reaction mixturewas cooled to room temp and the solvent was evaporated in vacuo tofurnish yellow viscous oil. This crude material was purified using aflash column packed with silica gel and eluted with 10% EtOAc/hexane toafford desired product as yellow oil (19.9 g, 64% yield). Anal. calcd.for C₁₄H₁₄Cl₂N₂O₂: C, 53.69; H, 4.52; N,8.94. Found: C, 53.33; H, 4.45;N, 8.56.

Part C: Ethyl 3-amino-2-(2,4-dichlorophenyl)-5-methylpyrrolecarboxylate: The compound of part B of Example 2 (19.9 g, 0.0635 moles)in ethanol (200 mL) was added dropwise 21% NaOEt in ethanol (22.6 mL,0.07 moles) at room temperature. The reaction mixture was furtherstirred at room temperature for 16 h. TLC (eluent 2% MeOH/CH₂Cl₂)revealed absence of starting material (Rf=0.86) and a new spot(Rf=0.33)was noticed. The reaction mixture was diluted with water (300 mL), solidseparated was filtered, washed with water and dried in a vacuum oven at60° C. for 2 days to afford orange yellow solid (15.7 g, mp 150-151° C.,79% yield). Anal. calcd. for C₁₄H₁₄Cl₂N₂O₂: C, 53.69; H, 4.52; N,8.94.Found: C, 53.44; H, 4.25; N, 9.04.

Part D:7-(2,4-Dichlorophenyl)-2,5-diimethyl-pyrrolo[3,4-d]pyrimidine-4(3H)-one:The compound of part C of Example 2 (10.0 g, 0.0322 mol) was dissolvedin dioxane (100.0 mL) and acetonitrile (100.0 mL) and cooled to 15° C.under nitrogen. Dry HCl (gas) was passed through a syringe needle intothe reaction mixture over a period of 30 mins. Stopped bubbling HCl gasat this stage and allowed to stir at room temperature for 20 h. Thesolvent was removed under vacuum, residue was dissolved in water (150mL) and basified using 28% ammonium hydroxide to pH 9 to 10. Filteredthe solid, washed with water, and dried to afford crude desired product.The crude was treated with hot 2-propanol (150 mL) and filtered theinsoluble white solid (7.35 g, mp >260° C., 74% yield). Anal. calcd. forC₁₄H₁₁Cl₂N₃O: C, 54.57; H, 3.61; N,13.64. Found: C, 54.20; H, 3.69; N,13.42.

Part E:4-Chloro-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidine:The compound of part D of Example 2 (1.0 g, 3.3 mmol) was treated withPOCl₃ (7.6 mL) and N,N-diisopropylethylamine (0.66 g, 5.1 mmol). Thereaction mixture was heated at 90° C. for 20 h. Excess POCl₃ was removedunder vacuum and then quenched with ice (50 g). The reaction mixture wasthen extracted extracted with EtOAc (3×50 mL), washed the combinedorganic layers with brine (50 mL) and dried with anhydrous magnesiumsulfate. The dried extract was filtered and concentrated in vacuum toafford a residue. The residue was purified by flash columnchromatography on a silica gel using dichloromethane as eluent to affordcorresponding chloro derivative (540 mg, mp 189-190° C., 51% yield) as ayellow solid. Anal. calcd. for C₁₄H₁₀Cl₃N₃: C, 51.48; H, 3.10; N,12.87.Found: C, 51.45; H, 3.08; N, 12.79.

Part F:4-[Bis[2-methoxyethyl)amino]-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidinehydrochloride salt: The compound of part E of Example 2 (0.1 g, 0.306mmol) in dichloromethane (10.0 mL) was treated with bis(2-methoxyethyl)amine (123 mg, 0.92 mmol, 3 eq.) and refluxed for 3days. The reaction mixture was concentrated in vacuum and recrystallizedthe solid to afford desired product as a yellow solid (HCl salt, 67 mg,mp 235-236° C., 48% yield). Anal. calcd. for C₂₀H₂₅Cl₃N₄O₂: C, 52.08; H,5.46; N,11.85. Found: C, 52.24; H, 5.48; N, 12.18.

Example 34-(N,N-Diethylamino)-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidine

The compound of part E of Example 2 (0.158 g, 0.48 mmol) indichloromethane (10.0 mL) was treated with N,N-diethylamine (180 mg, 2.4mmol, 5 eq.) and refluxed for 3 days. The reaction mixture wasconcentrated in vacuum and purified by flash column chromatography toafford desired product as light brown solid (15 mg, mp 146-148° C.). MScalcd. for C₁₈H₂₀Cl₂N₄:363.29. Found:363 (M+).

Example 44-(N-Cyclopropylmethyl-N-propylamino)-7-(2,4-dichlorophenyl)-2,5-dimethyl-pyrrolo[3,4-d]pyrimidine

The compound of part E of Example 2 (0.163 g, 0.5 mmol) in ethanol (10.0mL) was treated with N-propylcyclopropylmethyl-amine (113 mg, 1.0 mmol,2 eq.) and refluxed for 20 h. The reaction mixture was concentrated invacuum and purified by flash column chromatography to afford desiredproduct as yellow solid (59 mg, mp 66-68° C., 29% yield. MS calcd. forC₂₁H₂₄Cl₂N₄:403.36. Found:403 (M+).

Example 54-(N-Butyl-N-ethylamino)-7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine

The compound of part E of Example 2 (0.163 g, 0.5 mmol) in ethanol (10.0mL) was treated with N-ethylbutylamine (101 mg, 1.0 mmol, 2 eq.) andrefluxed for 22 h. The reaction mixture was concentrated in vacuum andpurified by flash column chromatography to afford desired product aspale yellow solid (120 mg, mp 94-95° C., 62% yield)

HRMS calcd. for C₂₀H₂₅Cl₂N₄:391.1456. Found: 391.1456 (M+H)

Example 64-[Bis(cyclopropylmethyl)amino]7-(2,4-dichlorophenyl)-2,5-dimethylpyrrolo[3,4-d]pyrimidine

The compound of part E of Example 2 (0.140 g, 0.43 mmol) in ethanol(10.0 mL) was treated with bis(cyclopropyl)methylamine hydrochloride(156 mg, 1.1 mmol) and Hunig's base (0.139 g, 1.1 mmol). The reactionmixture was refluxed for 22 h. The reaction mixture was concentrated invacuum and crude was treated with EtOAc (2.0 mL). The insoluble paleyellow solid was filtered to afford desired product as a HCl salt (173mg, mp 218-222° C., 92% yield). HRMS calcd. for C₂₁H₂₃Cl₂N₄:401.1300.Found: 401.1299 (M+H).

Example 77-(2,4-Dichloro-phenyl)-4-(1-ethyl-propylamino)-2,5-dimethyl-5H-pyrrolo[3,4-d]pyrimidin-5-ol

The compound of part E of Example 2 (0.065 g, 0.2 mmol) was treated with3-pentylamine (0.5 g, 5.8 mmol) and refluxed for 24 h. The reactionmixture was dissolved in dichloromethane (10 mL), washed with water (10mL) and dried. The dried organic extract was concentrated in vacuum andpurified by recrystallization from diethyl ether to afford tertiaryalcohol derivative as a white solid (17 mg, mp 184-185° C., 22% yield).NMR (CDCl₃) 0.95-1.0 (2 t, 6H, 2×CH₃), 1.4-1.75 (m, 4H, 2×CH₂), 1.8 (S,3H, CH₃), 2.55 (s, 3H, CH₃), 3.55 (bs, 1H, OH), 4.2-4.25 (m, 1H, CH),4.95 (d, 1H, NH), 7.4 (d, 1H, aromatic H), 7.55 (S, 1H, aromatic H),7.7-7.5 (d, 1H, aromatic H). MS calcd. for C₁₉H₂₂Cl₂N₄O: 393.32.Found:393 (M+).

Utility CRF—R1 Receptor Binding Assay for the Evaluation of BiologicalActivity

The following is a description of the isolation of cell membranescontaining cloned human CRF—R1 receptors for use in the standard bindingassay as well as a description of the assay itself.

Messenger RNA was isolated from human hippocampus. The mRNA was reversetranscribed using oligo (dt) 12-18 and the coding region was amplifiedby PCR from start to stop codons. The resulting PCR fragment was clonedinto the EcoRV site of pGEMV, from whence the insert was reclaimed usingXhoI+XbaI and cloned into the XhoI+XbaI sites of vector pm3ar (whichcontains a CMV promoter, the SV40 ‘t’ splice and early poly A signals,an Epstein-Barr viral origin of replication, and a hygromycin selectablemarker). The resulting expression vector, called phchCRFR wastransfected in 293EBNA cells and cells retaining the episome wereselected in the presence of 400 μM hygromycin. Cells surviving 4 weeksof selection in hygromycin were pooled, adapted to growth in suspensionand used to generate membranes for the binding assay described below.Individual aliquots containing approximately 1×10⁸ of the suspendedcells were then centrifuged to form a pellet and frozen.

For the binding assay a frozen pellet described above containing 293EBNAcells transfected with hCRFR1 receptors is homogenized in 10 ml of icecold tissue buffer (50 mM HEPES buffer pH 7.0, containing 10 mM MgCl₂, 2mM EGTA, 1 μg/l aprotinin, 1 μg/ml leupeptin and 1 μg/ml pepstatin). Thehomogenate is centrifuged at 40,000×g for 12 min and the resultingpellet rehomogenized in 10 ml of tissue buffer. After anothercentrifugation at 40,000×g for 12 min, the pellet is resuspended to aprotein concentration of 360 μg/ml to be used in the assay.

Binding assays are performed in 96 well plates; each well having a 300μl capacity. To each well is added 50 μl of test drug dilutions (finalconcentration of drugs range from 10-¹⁰-10-⁵ M), 100 μl of¹²⁵I-ovine-CRF (¹²⁵I-o-CRF) (final concentration 150 pM) and 150 μl ofthe cell homogenate described above. Plates are then allowed to incubateat room temperature for 2 hours before filtering the incubate over GF/Ffilters (presoaked with 0.3% polyethyleneimine) using an appropriatecell harvester. Filters are rinsed 2 times with ice cold assay bufferbefore removing individual filters and assessing them for radioactivityon a gamma counter.

Curves of the inhibition of ¹²⁵I-o-CRF binding to cell membranes atvarious dilutions of test drug are analyzed by the iterative curvefitting program LIGAND [P. J. Munson and D. Rodbard, Anal. Biochem.107:220 (1980), which provides Ki values for inhibition which are thenused to assess biological activity.

A compound is considered to be active if it has a K_(i) value of lessthan about 10000 nM for the inhibition of CRF.

Inhibition of CRF-Stimulated Adenylate Cyclase Activity

Inhibition of CRF-stimulated adenylate cyclase activity can be performedas described by G. Battaglia et al. Synapse 1:572 (1987). Briefly,assays are carried out at 37° C. for 10 min in 200 ml of buffercontaining 100 mM Tris-HCl (pH 7.4 at 37° C.), 10 mM MgCl₂, 0.4 mM EGTA,0.1% BSA, 1 mM isobutylmethylxanthine (IBMX), 250 units/mlphosphocreatine kinase, 5 mM creatine phosphate, 100 mM guanosine5′-triphosphate, 100 nM OCRF, antagonist peptides (concentration range10⁻⁹ to ₁₀ ⁻⁶ m) and 0.8 mg original wet weight tissue (approximately40-60 mg protein). Reactions are initiated by the addition of 1 mMATP/³²P]ATP (approximately 2-4 mCi/tube) and terminated by the additionof 100 ml of 50 mM Tris-HCL, 45 mM ATP and 2% sodium dodecyl sulfate. Inorder to monitor the recovery of cAMP, 1 μl of [³H]cAMP (approximately40,000 dpm) is added to each tube prior to separation. The separation of[³²P]cAMP from [³²P]ATP is performed by sequential elution over Dowexand alumina columns.

In Vivo Biological Assay

The in vivo activity of the compounds of the present invention can beassessed using any one of the biological assays available and acceptedwithin the art. Illustrative of these tests include the Acoustic StartleAssay, the Stair Climbing Test, and the Chronic Administration Assay.These and other models useful for the testing of compounds of thepresent invention have been outlined in C. W. Berridge and A. J. DunnBrain Research Reviews 15:71 (1990).

Compounds may be tested in any species of rodent or small mammal.

Compounds of this invention have utility in the treatment of inbalancesassociated with abnormal levels of corticotropin releasing factor inpatients suffering from depression, affective disorders, and/or anxiety.

Compounds of this invention can be administered to treat theseabnormalities by means that produce contact of the active agent with theagent's site of action in the body of a mammal. The compounds can beadministered by any conventional means available for use in conjunctionwith pharmaceuticals either as individual therapeutic agent or incombination of therapeutic agents. They can be administered alone, butwill generally be administered with a pharmaceutical carrier selected onthe basis of the chosen route of administration and standardpharmaceutical practice.

The dosage administered will vary depending on the use and known factorssuch as pharmacodynamic character of the particular agent, and its modeand route of administration; the recipient's age, weight, and health;nature and extent of symptoms; kind of concurrent treatment; frequencyof treatment; and desired effect. For use in the treatment of saiddiseases or conditions, the compounds of this invention can be orallyadministered daily at a dosage of the active ingredient of 0.002 to 200mg/kg of body weight. Ordinarily, a dose of 0.01 to 10 mg/kg in divideddoses one to four times a day, or in sustained release formulation willbe effective in obtaining the desired pharmacological effect.

Dosage forms (compositions) suitable for administration contain fromabout 1 mg to about 100 mg of active ingredient per unit. In thesepharmaceutical compositions, the active ingredient will ordinarily bepresent in an amount of about 0.5 to 95% by weight based on the totalweight of the composition.

The active ingredient can be administered orally is solid dosage forms,such as capsules, tablets and powders; or in liquid forms such aselixirs, syrups, and/or suspensions. The compounds of this invention canalso be administered parenterally in sterile liquid dose formulations.

Gelatin capsules can be used to contain the active ingredient and asuitable carrier such as but not limited to lactose, starch, magnesiumstearate, steric acid, or cellulose derivatives. Similar diluents can beused to make compressed tablets. Both tablets and capsules can bemanufactured as sustained release products to provide for continuousrelease of medication over a period of time. Compressed tablets can besugar-coated or film-coated to mask any unpleasant taste, or used toprotect the active ingredients from the atmosphere, or to allowselective disintegration of the tablet in the gastrointestinal tract.

Liquid dose forms for oral administration can contain coloring orflavoring agents to increase patient acceptance.

In general, water, pharmaceutically acceptable oils, saline, aqueousdextrose (glucose), and related sugar solutions and glycols, such aspropylene glycol or polyethylene glycol, are suitable carriers forparenteral solutions. Solutions for parenteral administration preferablycontain a water soluble salt of the active ingredient, suitablestabilizing agents, and if necessary, butter substances. Antioxidizingagents, such as sodium bisulfite, sodium sulfite, or ascorbic acid,either alone or in combination, are suitable stabilizing agents. Alsoused are citric acid and its salts, and EDTA. In addition, parenteralsolutions can contain preservatives such as benzalkonium chloride,methyl- or propyl-paraben, and chlorobutanol.

Suitable pharmaceutical carriers are described in “Remington'sPharmaceutical Sciences”, A. Osol, a standard reference in the field.

Useful pharmaceutical dosage-forms for administration of the compoundsof this invention can be illustrated as follows:

Capsules

A large number of units capsules are prepared by filling standardtwo-piece hard gelatin capsules each with 100 mg of powdered activeingredient, 150 mg lactose, 50 mg cellulose, and 6 mg magnesiumstearate.

Soft Gelatin Capsules

A mixture of active ingredient in a digestible oil such as soybean,cottonseed oil, or olive oil is prepared and injected by means of apositive displacement was pumped into gelatin to form soft gelatincapsules containing 100 mg of the active ingredient. The capsules werewashed and dried.

Tablets

A large number of tablets are prepared by conventional procedures sothat the dosage unit was 100 mg active ingredient, 0.2 mg of colloidalsilicon dioxide, 5 mg of magnesium stearate, 275 mg of microcrystallinecellulose, 11 mg of starch, and 98.8 mg lactose. Appropriate coatingsmay be applied to increase palatability or delayed adsorption.

The compounds of this invention may also be used as reagents orstandards in the biochemical study of neurological function,dysfunction, and disease.

Although the present invention has been described and exemplified interms of certain preferred embodiments, other embodiments will beapparent to those skilled in the art. The invention is, therefore, notlimited to the particular embodiments described and exemplified, but iscapable of modification or variation without departing from the spiritof the invention, the full scope of which is delineated by the appendedclaims.

What is claimed is:
 1. A compound of Formula (II),

wherein: D is Ar¹ or heteroaryl, each optionally substituted with 1 to 5R⁵ groups and each is attached to an unsaturated carbon atom; R¹ is H,Ar², heteroaryl, heterocyclyl, or carbocyclyl; or C₁-C₆ alkyl, C₂-C₄alkenyl, or C₂-C₄ alkynyl, each optionally substituted with 1 to 3substituents independently selected at each occurrence from C₁-C₄ alkyl,Ar², heteroaryl, heterocyclyl, carbocyclyl, OR¹², F, Cl, Br, I, CF₃, andNO₂; R³ is H, Ar², heteroaryl, heterocyclyl, or carbocyclyl; or C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, or C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, Ar², heteroaryl, heterocyclyl,carbocyclyl, NO₂, F, Cl, Br, I, NR⁹COR⁷, NR⁹CO₂R⁷, OR⁷, CONR¹OR¹¹, andCO(NOR¹²)R⁷; R⁴ is NR¹¹R¹⁰; R⁵ is independently selected at eachoccurrence from: NO₂, F, Cl, Br, I, CN, NR¹OR¹¹, NR⁹COR¹³, NR⁹CO₂R⁷.COR¹³, R¹³, OR¹², CONR¹⁰R¹¹, CO(NOR⁹)R¹⁰, CO₂R¹², and S(O)_(n)R¹⁴; orC₁-C₆ alkyl, C₂-C₁₀ alkenyl, and C₂-C₁₀ alkynyl, each optionallysubstituted with 1 to 3 substituents independently selected at eachoccurrence from C₁-C₄ alkyl, C₃-C₆ cycloalkyl, CF₃, NO₂, F, Cl, Br, I,CN, NR⁶R⁷, NR⁹COR⁷, NR⁹CO₂R⁷, COR⁷, OR⁷, CONR⁶R⁷, CO₂R⁷, CO(NOR⁹)R⁷, andS(O)_(n)R⁷; R⁶ is independently selected at each occurrence from: H,—CH₂Ar², Ar², heteroaryl, heterocyclyl, and carbocyclyl; or C₁-C₁₀alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, and C₁-C₄ haloalkyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₆ alkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I,OR¹², NO₂, S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹⁴, NR⁹COR¹³, N(COR¹³)₂,NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰, CONR¹¹R¹⁰, Ar², heteroaryl,heterocyclyl, and carbocyclyl; R⁷ is independently selected at eachoccurrence from: H, —CH₂Ar², Ar², heteroaryl, heterocyclyl, andcarbocyclyl; or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, and C₁-C₄haloalkyl, each optionally substituted with 1 to 3 substituentsindependently selected at each occurrence from C₁-C₆ alkyl, C₁-C₄haloalkyl, CN, F, Cl, Br, I, OR¹², NO₂, S(O)_(n)R¹⁴, COR¹³, CO₂R¹²,OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹¹R¹⁰,CONR¹¹R¹⁰, Ar², heteroaryl, heterocyclyl, and carbocyclyl; Ar¹ is phenylor naphthyl; Ar² is phenyl or naphthyl, each optionally substituted with1 to 5 substituents independently selected at each occurrence from C₁-C₆alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, CN, F, Cl, Br, I, OR¹², NO₂,S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰,NR⁹CO₂R¹², NR¹¹R¹⁰, and CONR¹¹R¹⁰; heteroaryl is pyridyl, pyrimidinyl,triazinyl, furanyl, pyrrolyl, imidazolyl, pyranyl, quinolinyl,isoquinolinyl, thienyl, imidazolyl, thiazolyl, indolyl, indazolyl,pyrrolyl, oxazolyl, benzofuranyl, benzothienyl, benzothiazolyl,isoxazolyl, pyrazolyl, 2,3-dihydrobenzothienyl or2,3-dihydrobenzofuranyl; heterocyclyl is saturated or partiallysaturated heteroaryl, optionally substituted with 1 to 5 substituentsindependently selected at each occurrence from C₁-C₆ alkyl, C₃-C₆cycloalkyl, C₁-C4 haloalkyl, CN, F, Cl, Br, I, OR¹², NO₂, S(O)_(n)R¹⁴,COR¹³, CO₂R12, OC(O)R¹³, NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹²,NR¹⁰R¹¹, and CONR¹¹R¹⁰; carbocyclyl is saturated or partiallyunsaturated C₃-C₁₀ membered ring, optionally substituted with 1 to 5substituents independently selected at each occurrence from C₁-C₆ alkyl,CF₃, CN, F, Cl, Br, I, OR¹², SR¹⁰, S(O)_(n)R¹⁴, COR¹³, CO₂R¹², OC(O)R¹³,NR⁹COR¹³, N(COR¹³)₂, NR⁹CONR¹¹R¹⁰, NR⁹CO₂R¹², NR¹⁰R¹¹, and CONR¹¹R¹⁰; R⁹is independently selected at each occurrence from H, C₁-C₄ alkyl, andC₃-C₆ cycloalkyl; R¹⁰ is H, heterocyclyl, or carbocycle; or C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, Ar², or heteroaryl, each optionallysubstituted with 1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²; R¹¹ is H,heterocyclyl, or carbocycle; or C₁-C₆ alkyl, C₂-C₆ alkenyl, and C₂-C₆alkynyl, Ar², or heteroaryl, each optionally substituted with 1-3 C₁-C₆alkyl, C₃-C₆ cycloalkyl, F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²;alternatively, R¹⁰ and R¹¹ can combine to form piperidine, pyrrolidine,piperazine, N-methylpiperazine, morpholine or thiomorpholine, eachoptionally substituted with 1-3 C₁-C₄ alkyl groups; R¹² is independentlyselected at each occurrence from H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, andC₁-C₄ haloalkyl; R¹³ is selected from H, C₁-C₄ alkyl, C₁-C₄ haloalkyl,C₁-C₄ alkoxy C₁-C₆ alkyl, C₃-C₆ cycloalkyl, and C₃-C₆ cycloalkyl C₁-C₆alkyl R¹⁴ is independently selected at each occurrence from C₁-C₆ alkyl,C₃-C₆ alkyl, and phenyl, each subsituted by 1-3 C₁-C₄ alkyl, C₁-C₄haloalkyl or OR¹⁵; and R¹⁵ is independently selected at each occurrencefrom H, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, and C₁-C₄ haloalkyl.
 2. Acompound of claim 1 of Formula (IIa),

wherein: R¹ is H; or C₁-C₆ alkyl, C₂-C₄ alkenyl, or C₂-C₄ alkynyl, eachoptionally substituted with 1 to 3 substituents independently selectedat each occurrence from C₁-C₄ alkyl, OR¹², F, Cl, Br, I, CF₃, and NO₂;R³ is H; or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, or C₁-C₄haloalkyl, each optionally substituted with 1 to 3 substituentsindependently selected at each occurrence from C₁-C₄ alkyl, NO₂, F, Cl,Br, I, NR⁹COR⁷, NR⁹CO₂R⁷, OR⁷, CONR¹⁰R¹¹, or CO(NOR¹²)R⁷; R⁴ is NR¹¹R¹⁰;R⁵ is independently selected at each occurrence from: NO₂, F, Cl, Br, I,CN, and R¹³; R⁷ is independently selected at each occurrence from: H,—CH₂Ar², and Ar²; or C₁-C₁₀ alkyl, C₂-C₁₀ alkenyl, C₂-C₁₀ alkynyl, andC₁-C₄ haloalkyl, each optionally substituted with 1 to 3 substituentsindependently selected at each occurrence from C₁-C₆ alkyl, C₁-C₄haloalkyl, CN, F, Cl, Br, I, NO₂, and OR¹²; Ar² is phenyl optionallysubstituted with 1 to 5 substituents independently selected at eachoccurrence from C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₄ haloalkyl, CN, F,Cl, Br, I, OR¹², and NO₂; R⁹ is independently selected at eachoccurrence from H, C₁-C₄ alkyl, and C₃-C₆ cycloalkyl; R¹⁰ isindependently selected at each occurrence from H, C₁-C₆ alkyl, C₂-C₆alkenyl, and C₂-C₆ alkynyl, each optionally substituted with 1-3 F, Cl,Br, I, NO₂, CF₃, or OR¹²; R¹¹ is independently selected at eachoccurrence from H, C₁-C₆ alkyl, C₂-C₆ alkenyl, and C₂-C₆ alkynyl, eachoptionally substituted with 1-3 C₁-C₆ alkyl, C₃-C₆ cycloalkyl, F, Cl,Br, I, NO₂, CF₃, or OR¹²; alternatively, R¹⁰ and R¹¹ can combine to formpiperidine, pyrrolidine, piperazine, N-methylpiperazine, morpholine orthiomorpholine, each optionally substituted with 1-3 C₁-C₄ alkyl groups;R¹² is independently selected at each occurrence from H, C₁-C₆ alkyl,C₃-C₆ cycloalkyl, and C₁-C₄ haloalkyl; R¹³ is selected from H, C₁-C₄alkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy C₁-C₆ alkyl, C₃-C₆ cycloalkyl, andC₃-C₆ cycloalkyl C₁-C₆ alkyl.
 3. A compound of claim 2 wherein: R⁵ isindependently selected at each occurrence from: NO₂, F, Cl, Br, I, CN,and R¹³; R⁴ is NR¹⁰R¹¹; R¹⁰ is H; or methyl, ethyl, propyl, butyl,ethene, propene, butene, or propargyl, each optionally substituted with1-3 F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²; R¹¹ is H; or methyl, ethyl,propyl, butyl, ethene, or propene, each optionally substituted with 1-2methyl, ethyl, propyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,F, Cl, Br, I, NO₂, CF₃, CN, or OR¹²; alternatively, R¹⁰ and R¹¹ cancombine to form piperidine, pyrrolidine, piperazine, N-methylpiperazine,morpholine or thiomorpholine, each optionally substituted with 1-3methyl, ethyl, or propyl groups.
 4. A compound of claim 1 that is:7-(2,4-Dichloro-phenyl)-4-(1-ethyl-propylamino)-2,5-dimethyl-5H-pyrrolo[3,4-d]pyrimidin-5-ol.5. A pharmaceutical composition comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound accordingto claim
 1. 6. A method of treating affective disorder, anxiety, ordepression in a mammal comprising administering to the mammal a dose ofa composition according to claim 5.